Method for lowering blood pressure in pre-hypertensive individuals and/or individuals with metabolic syndrome

ABSTRACT

The present invention relates to a method of treating Metabolic Syndrome and/or the conditions that comprise Metabolic Syndrome by administering to a mammal, including a human, a dietary supplement comprising grape extract. This invention also relates to a method of treating pre-hypertension by administering to a mammal, including a human, a dietary supplement comprising grape extract.

This application claims priority to U.S. Provisional application Ser.No. 60/721,720, filed on Sep. 28, 2005, which is incorporated byreference in its entirety.

FIELD OF THE INVENTION

This invention relates to a method of preventing and/or treatingMetabolic Syndrome and/or the conditions that comprise MetabolicSyndrome. This invention also relates to a method of treating and/orpreventing pre-hypertension. These methods involve administering to amammal, including a human, a dietary supplement comprising grape extracteffective for treating pre-hypertensive individuals, or individuals withMetabolic Syndrome, and/or the conditions that comprise MetabolicSyndrome.

BACKGROUND

Blood pressure is the force exerted by the bloodstream on the arterywalls. Normal blood pressure is considered to be 120 systolic and 80diastolic. Hypertension refers to a disorder that is characterized by anelevation of the systolic blood pressure to 140 and above and/or anelevation of the diastolic blood pressure to 90 and above. Withhypertension, there is either an increase in the total peripheralvascular resistance such as is due to vasoconstriction, or an increasein cardiac output, or both. These conditions produce an elevation inblood pressure because blood pressure is equal to flow times resistance.There are many factors that can contribute to high blood pressureincluding stress, diet and lifestyle, as well as kidney complaints,hormonal disturbances and circulatory disorders. An untreatedhypertensive patient is at great risk of developing left ventricularfailure, myocardial infarction, cerebral hemorrhage or renal failure.Hypertension is also a risk factor for stroke and coronaryatherosclerosis. Presently, hypertensive patients are treated with drugtherapy that includes the use of diuretics, beta-blockers, ACEinhibitors, angiotensin antagonists, calcium channel blockers,alpha-blockers, alpha-beta-blockers, nervous system inhibitors, andvasodilators.

Pre-hypertensive individuals are classified as individuals that havesystolic pressure between 120 and 139 mmHg or have diastolic pressurebetween 81 and 89 mmHG. This classification is based on the SeventhReport of the Joint National Committee on Prevention, Detection,Evaluation, and Treatment of High Blood Pressure (JNC 7), page 87, NIHPublication No. 04-5230. Pre-hypertensive individuals are not typicallytreated with drug therapy, but rather are given suggestions for ahealthy lifestyle. These suggestions include maintaining a healthyweight; being physically active; following a healthy eating plan thatemphasizes fruits, vegetables, and low fat dairy foods; choosing andpreparing foods with less sodium; and drinking alcoholic beverages inmoderation if at all. Adopting healthy lifestyle habits is usually aneffective first step in both preventing and controlling abnormal bloodpressure.

Many patients with hypertension are insulin resistant. Insulinstimulates glucose uptake into tissues, and its ability to do so variesgreatly among individuals. With insulin resistance, tissues have adiminished ability to respond to the action of insulin. To compensatefor resistance, the pancreas secretes more insulin. Insulin-resistantindividuals, therefore, have high plasma insulin levels. There isevidence that abnormal blood pressure is linked to the degree of insulinresistance. The precise way in which insulin resistance develops isunclear, although genetics, diet and level of physical activity arebelieved to play a role.

“Metabolic Syndrome,” also called “Syndrome X,” the “Insulin ResistanceSyndrome,” or the “Deadly Quartet,” is characterized by an accumulationof risk factors for cardiovascular disease, stroke and/or diabetesmellitus type II. Metabolic Syndrome may be caused by an overproductionof cortisol, a stress hormone, which causes an accumulation of fatinside the abdominal cavity and insulin resistance. Drug therapy is notcurrently recommended for individuals with Metabolic Syndrome. The riskfactors that characterize Metabolic Syndrome include an increased amountof adipose tissue inside the abdominal cavity (abdominal obesity),insulin resistance with increased risk of developing diabetes,hyperinsulinemia, high levels of blood fats, increased blood pressure,and elevated serum lipids. The National Cholesterol Education AdultTreatment Panel (ATP III) defined Metabolic Syndrome as individualshaving at least three of the following risk factors:

Risk Factor Defining Level Abdominal obesity, given as waistcircumference*^(†) Men >102 cm (>40 in) Women >88 cm (>35 in)Triglycerides ≧150 mg/dL HDL cholesterol Men <40 mg/DI Women <50 mg/dLBlood pressure ≧130/≧85 mm Hg Fasting glucose ≧110 mg/dL^(‡) *Overweightand obesity are associated with insulin resistance and MetabolicSyndrome. The presence of abdominal obesity, however, is more highlycorrelated with the metabolic risk factors than is an elevated BMI.Therefore, the simple measure of waist circumference is recommended toidentify the body weight component of Metabolic Syndrome. ^(†)Some malepatients can develop multiple metabolic risk factors when the waistcircumference is only marginally increased, e.g., 94 to 102 cm (37 to 39in). Such patients may have a strong genetic contribution to insulinresistance. They should benefit from changes in life habits, similarlyto men with categorical increases in waist circumference. ^(‡)TheAmerican Diabetes Association has recently established a cut-off pointof ≧100 mg/dL, above which individuals have either pre-diabetes(impaired fasting glucose) or diabetes. This new cut-off point should beapplicable for identifying the lower boundary to define an elevatedglucose as one criterion for Metabolic Syndrome.

The World Health Organization, however, defines Metabolic Syndrome asindividuals with diabetes/insulin resistance and at least two of thefollowing risk factors: high waist-to -hip ratio; high triglycerides orlow HDL cholesterol; high blood pressure; and a high urinary albuminexcretion rate.

Conditions related to Metabolic Syndrome include diabetes mellitus typeII, dyslipoproteinemia, myocardial infarction, stroke and otherarteriosclerotic diseases, as well as the risk factors for thesediseases, including insulin resistance in general, abdominal obesitycaused by accumulation of intra-abdominal fat, elevated blood serumlipids and glucose, raised diastolic and/or systolic blood pressure, andhypertension.

Treatment of pre-hypertensive individuals and individuals with MetabolicSyndrome are encouraged to adopt a healthy lifestyle, which includesmaintaining a healthy weight; being physically active; and following ahealthy eating plan. Given that drug therapy is not recommended to theseindividuals, there is a need for a dietary supplement comprising grapeextract that these individuals can use as adjunctive therapy, which iseffective in lowering blood pressure and does not increase insulinresistance.

Grape seeds contain about 5-8% by weight flavonoids. Flavonoidsconstitute an important group of dietary polyphenolic compounds that arewidely distributed in plants. More than 4000 chemically uniqueflavonoids have been identified in plant sources, such as fruits,vegetables, legumes, nuts, seeds, herbs, spices, flowers, as well as inbeverages such as tea, cocoa, beer, wine, and grape juice.

The terminology of flavonoids with respect to grape seeds refers tomonomeric flavan-3-ols, specifically (+)-catechin, (−)-epicatechin, and(−)-epicatechin 3-gallate. Two or more flavan-3-ol monomers chemicallylinked are called proanthocyanidins or oligomeric proanthocyanidins(“OPCs”), which includes procyanidins and prodelphinidins. OPCscontaining two monomers are called dimers, three monomers are calledtrimers, four monomers are called tetramers, five monomers are calledpentamers, etc. Operationally, the oligomers have chain lengths of 2 to7 (dimers to heptamers); whereas polymers represent components withchain lengths greater than 7. After considerable discussion, it was theconsensus of the Grape Seed Method Evaluation Committee (through theNational Nutritional Foods Association) to define OPCs as allproanthocyanidins containing two or more monomers, including polymers orcondensed tannins. Thus, oligomers in grape extracts include, forinstance, dimers and trimers, and there is evidence that the polymerscan have as many as sixteen units.

Below is a typical structure of a proanthocyanidin, showingepicatechin-gallate extension units and terminal units. The extensionunits are represented, for instance, by the epicatechin (2) andepigallocatechin (3) linking groups. Whereas, a terminal unit isrepresented by the epicatechin gallate (4) group.

In order for polyphenolic compounds to be used commercially as a grapeextract, these compounds have to be separated from grapes in a moreconcentrated form. The general process in which the polyphenoliccompounds are extracted, purified and concentrated from whole grapes,grape pomace and grape seeds is disclosed in U.S. Pat. No. 6,544,581,which is incorporated herein by reference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the relationship between the baseline blood pressure andthe decrease in systolic pressure for individual with Metabolic Syndromebeing treated with the grape extract used in the present invention.

FIG. 2 shows the relationship between the baseline blood pressure andthe decrease in diastolic pressure for individual with MetabolicSyndrome being treated with the grape extract used in the presentinvention.

FIG. 3 shows the change in oxidized LDL concentration in individualswith Metabolic Syndrome being treated with the grape extract used in thepresent invention.

FIG. 4 shows the relationship between the change in oxidized LDLconcentration and the baseline concentration of oxidized LDL inindividuals given 300 mg of the grape extract used in the presentinvention.

DETAILED DESCRIPTION

The present invention provides a method of preventing and/or treatingMetabolic Syndrome and/or the conditions that comprise MetabolicSyndrome. This method includes administering to a mammal, including ahuman, in need of such treatment a dietary supplement comprising aneffective amount of grape extract. In particular, the grape extract usedin the present method is effective to lower blood pressure and reduceoxidized LDL cholesterol in individual with Metabolic Syndrome.

The present invention also provides a method of treating and/orpreventing Metabolic Syndrome in a mammal, comprising administering adietary supplement comprising grape extract having about 5-15% monomers,about 5-20% dimers, about 3-10% trimers, about 2-10% tetramers, andabout 2-10% pentamers by weight. The total amount of low molecularweight phenolic compounds including monomers, dimers, trimers,tetramers, and pentamers is between about 25-50% by weight, preferablybetween about 25-40% by weight, more preferably between about 30-40% byweight, and more preferably between about 25-35% by weight. The totalamount of phenolic compounds is about 80% by weight or more, andpreferably about 90% by weight or more.

The present invention also provides a method of preventing and/ortreating pre-hypertension. This method includes administering to amammal, including a human, in need of such treatment a dietarysupplement comprising an effective amount of grape extract. Inparticular, the grape extract used in the present method is effective tolower blood pressure in pre-hypertensive individuals.

The present invention also provides a method of treating and/orpreventing pre-hypertension in a mammal, comprising administering adietary supplement comprising grape extract having about 5-15% monomers,about 5-20% dimers, about 3-10% trimers, about 2-10% tetramers, andabout 2-10% pentamers by weight. The total amount of low molecularweight phenolic compounds including monomers, dimers, trimers,tetramers, and pentamers is between about 25-50% by weight, preferablybetween about 25-40% by weight, more preferably between about 30-40% byweight, and more preferably between about 25-35% by weight. The totalamount of phenolic compounds is about 80% by weight or more, andpreferably about 90% by weight or more.

The term “effective amount” means an amount of grape extract sufficientto decrease either or both systolic and/or diastolic blood pressure inpre-hypertensive individuals or those with Metabolic Syndrome by atleast about 2%, preferably by at least about 5%, and more preferably byat least about 8%, without having adverse effects such. as increasinginsulin resistance in the individual. An appropriate clinical end-point,however, is a systolic blood pressure below 120 mmHG in pre-hypertensiveindividuals or those with Metabolic Syndrome. It has also been suggestedthat the grape extract used in the present invention will, in additionto lowering blood pressure, reduce oxidized LDL cholesterol inindividuals with Metabolic Syndrome. Increased LDL cholesterol is arecognized risk factor for atherosclerosis. There is strong evidencethat oxidatively modified LDL initiates the development of thispathological process. Thus, decreasing the concentration of oxidized LDLmay reduce and/or prevent atherosclerosis in individuals with MetabolicSyndrome.

The grape extract, to be used with the present invention, has a phenolicprofile, as determined by normal-phase high-performance liquidchromatography (“HPLC”), of about 5-15% monomers, about 5-20% dimers,about 4-10% trimers, about 2-10% tetramers, and about 2-10% pentamers byweight. The grape extract used in the present invention also comprisesabout 80% by weight or more total phenolic compounds, and preferablyabout 90% by weight or more, as determined by the Folin Ciocalteumethod. The grape extract used in the present invention also comprisesabout 2% by weight or less epicatechin-gallate terminal units, morepreferably about 1% by weight or less, as determined by reverse-phaseHPLC after thiolysis reaction. The grape extract used in the presentinvention also comprises about 12% by weight or less epicatechin-gallateextension units, preferably about 8% by weight or less, and morepreferably about 5% by weight or less, as determined by reverse-phaseHPLC after thiolysis reaction.

The grape extract used in the present invention may be produced bymodifying the hot water extraction process disclosed in U.S. Pat. No.6,544,581 as described below. In general, the hot water extractionprocess, as disclosed in the '581 patent, involves the following steps.In step (1), grape seeds, dry or fresh, may be heated with hot water fora time sufficient to extract most of the polyphenols. Temperatures of140-212° F. may be employed, preferably 160-212° F., more preferably180-212° F., yet more preferably 190-212° F., for a period of about 1-6hours. The time of heating may be varied in relation to the temperatureused. Generally, lower temperatures require longer extraction times. Instep (2), the crude grape seed-water extract may be separated from spentseeds by draining over metal screens. The extract may then be cooled andtreated with any suitable commercially available pectolytic enzyme, suchas Pectinex® Ultra SP-L manufactured by Novo Nordisk, at a concentrationof about 50-200 ppm to break down cell wall constituents. Preferably,the seed water extract may be enzyme-treated for a period of two hoursat a temperature of 80-120° F. Alternatively, the seed-water extract maybe enzyme-treated for 7-14 days or longer at about 40-50° F. In step(3), the resulting turbid seed extract may be acidified with an acid,preferably a mineral acid, more preferably with sulfuric acid, to a pHof approximately 1.5-2.5 and allowed to react from about one hour toabout two days. The acidified extract may be cooled for up to severalweeks to allow for macromolecules, including proteins and otherpolysaccharides, to settle. The cooled acidified extract may then befiltered using diatomaceous earth to yield a clarified seed extract.Other filter aids, such as perlite, may also be used.

Step (2) of the '581 patent may be modified by enzyme-treating theseed-water extract for a period of four to five days at a temperature ofabout 80-120° F. in order to produce the grape extract used in thepresent invention. While not intending to be bound to any theory, it isbelieved that the longer duration than used in the '581 patent at thespecified temperature range of this step accounts for the resultingnovel grape extract. The time of the enzyme treatment may be varied inrelation to the temperature used. Generally, lower temperatures requirelonger treatment times. Thus, the seed-water extract may beenzyme-treated for up to two weeks or longer at temperatures of about60-80° F.

Alternatively, the grape extract used in the present invention may beproduced by the following steps. After the extraction of step (1) orafter the pectinase treatment in step (2) of the '581 patent, theextract may be smeared on a bacteriological agar plate. Upon incubation,plural species of yeast, bacteria, and/or fungi may be present dependingupon the starting material. The live culture may be isolated as acocktail. Once isolated, the cocktail may be used in subsequentextraction and/or pectinase enzyme treatment steps. For instance, aseed-water extract may be enzyme treated with any suitable commerciallyavailable pectolytic enzyme and combined with the isolated cocktail ofyeast, bacteria, and/or fungi. The combined mixture may be allowed tostand for a period of about one to ten days, preferably about two tofive days, at a temperature of about 70-100° F. The time of the enzymetreatment may be varied in relation to the temperature used and theinoculum count. The resulting turbid seed extract may be acidified witha suitable acid as discussed above to a pH of 1.5-2.5 and allowed toreact for about one hour to about two days. The acidified extract may becooled and stored for several days to allow for flocculation of proteinsand polysaccharides. The cool acidified extract may then be filteredusing diatomaceous earth to yield a clarified seed extract, which may befurther processed according to the '581 patent to produce a purifiedgrape extract suitable for blood pressure reduction and reduction ofoxidized LDL.

The amount of gallic acid in grape extracts produced by the '581 processas compared with grape extracts produced by the process involving acocktail of yeast, bacteria, and/or fungi was analyzed by HPLC. Suchanalysis showed an increase from a value of between about 50-150 ppm ofgallic acid in the grape extracts of the '581 process to a value ofbetween about 400-1500 ppm of gallic acid in the grape extracts used inthe present invention using the cocktail. The increase in gallic acidindicates that the epicatechin-gallate terminal and extension units arede-esterfied from procyanidins. While not intending to be bound to anytheory, it is believed that the cocktail of yeast, bacteria, and/orfungi use the grape extract as a substrate for growth and producetannase enzymatic activity, which results in de-esterification ofprocyanidins and release of gallic acid. As such, using a cocktail oflive yeast, bacteria, and/or fungi produces the grape extract used inthe present invention having about 2% by weight or lessepicatechin-gallate terminal units, more preferably about 1% by weightor less, and about 12% by weight or less epicatechin-gallate extensionunits, preferably about 8% by weight or less, and more preferably about5% by weight or less.

In one embodiment, the grape extract of the present invention may beproduced by the following steps. After the extraction of step (1) orafter the pectinase treatment in step (2) of the '581 patent, anysuitable commercial fungal tannase enzyme, such as tannin acylhydrolase,E.C3.1.1.20, may be added at a concentration of between about 5-1000ppm. Depending on the concentration of the tannase enzyme used, themixture may be reacted for about one hour to about two days, preferablyone to two days, or until the terminal units are reduced to about 2% orless, preferably 1% or less, and the extension units are reduced toabout 8% or less, preferably about 5% or less. After a sufficientreaction time, the extract may be acidified to a pH of 1.5 to 2.5, whichallows flocculation of proteins and polysaccharides on cooler storagefrom 40-60° F. The extract may be filtered to clarify and processedfurther according to the '581 patent to produce a grape extract withcharacteristics for blood pressure reduction.

The grape extract used in the present invention may be formulated intodietary supplements, including capsules, tablets, powders, solutions,gels, suspensions, creams, pastes, gels, suppositories, transdermalpatches, and the like. These dietary supplements in, for instance,powder or solution form, may be added to nutraceuticals, foods and/orbeverages to form functional nutraceutical, food, and/or beverageproducts. The dietary supplements may be formulated as powders, forexample, for mixing with consumable liquids such as milk, juice, wateror consumable gels or syrups for mixing into other dietary liquids orfoods. The dietary supplements of this invention may be formulated withother foods or liquids to provide pre-measured supplemental foods, suchas single serving bars. Typical food products that may incorporate thegrape extract used in the present invention include dairy foods such asyogurt, cereals, breads, snack food products, fruit juices and othersoft drinks. Flavorings, binders, protein, complex carbohydrates,vitamins, minerals and the like may be added as needed. Preferably, thegrape extract is formulated for oral administration.

The dietary supplements used in the present invention are intended fordaily administration or as needed. The magnitude of a prophylactic ortherapeutic dose of the dietary supplement in pre-hypertensiveindividuals or those with Metabolic Syndrome will vary with the severityof the condition being treated and the route of administration. Thedose, and perhaps the dose frequency, will also vary according to theage, body weight, and response of the individual. In general, the totaldaily dose range, for the conditions described herein, is from about 50mg to about 1,000 mg by weight of grape extract administered in singleor divided doses orally, topically, or transdermally, preferably orally.A preferred oral daily dose range is from about 50 mg to about 500 mg byweight of the grape extract (i.e., excluding excipients and carriers),more preferably about 150 mg to about 300 mg. For example, capsules ortablets may be formulated in either 150 mg or 300 mg doses, whereasbeverages can be formulated with 50 mg grape extract. Such a regimen ofadministration is preferably maintained for at least one month, morepreferably six months or longer.

The dietary supplements used in the present invention may be formulatedin a conventional manner (e.g., wet or dry granulation), in admixturewith pharmaceutically acceptable carriers, excipients, vitamins,minerals and/or other nutrients. Representative carriers and excipientsinclude, but are not limited to, starches, sugars, microcrystallinecellulose, diluents, granulating agents, lubricants, binders,disintegrating agents, and the like, in the case of oral solidpreparations (such as powders, capsules, and tablets).

Any suitable route of administration may be employed to administer thedietary supplements of the invention to an individual. Suitable routesinclude, for example, oral, rectal, parenteral, intravenous, topical,transdermal, subcutaneous, and intramuscular. Although any suitableroute of administration may be employed for providing the individualwith an effective amount of the grape extract according to the methodsof the present invention, oral administration is preferred, includingsolid dosage forms such as tablets, capsules, or powders. It is alsopreferred that the grape extract is formulated for use in functionalnutraceutical, food, or beverage products.

The grape extract used in the present invention can also be combinedwith other active agents including but not limited to diuretics,beta-blockers, ACE inhibitors, angiotensin antagonists, calcium channelblockers, alpha-blockers, alpha-beta-blockers, nervous systeminhibitors, vasodilators, antioxidants.

1. Characterization of Grape Extracts

Recently, it was reported that the use of grape seed polyphenols doesnot reduce systolic blood pressure and actually increases systolic bloodpressure when combined with the use of vitamin C in hypertensiveindividuals. See Ward et al. “The combination of vitamin C andgrape-seed polyphenols increases blood pressure: a randomized,double-blind, placebo-controlled trial,” Journal of Hypertension 2005;23:427434. While not intending to be bound to any theory, it is believedthat the phenolic profile of grape extracts is important to theireffectiveness in reducing blood pressure. The grape extract evaluated inthe Ward study was Vinlife®, which has a phenolic profile of 50.6% totalphenolic compounds, as determined by Folin Ciocalteu method, 11.2%epicatechin-gallate terminal units, and 11.8% epicatechin-gallateextension units, as determined by reverse-phase HPLC after thiolysisreaction, and 7.3% monomers, 4.4% dimers, 2.0% trimers, 1.9% tetramers,and 1.1% pentamers, with a total monomers to pentamers of 16.7%, asdetermined by normal-phase HPLC.

Commercially available extracts of grape seed contain a variety ofmonomers and proanthocyanidins. The phenolic profile of somecommercially available extracts, as determined by reverse-phase HPLC, isdescribed in Table 1, and as determined by normal-phase HPLC, isdescribed in Table 2. From these analyses, the grape extract used in thepresent invention (currently manufactured by Polyphenolics, Inc. asMegaNatural®-BP) has three differentiating factors that distinguish itfrom other grape extracts:

-   -   1. High degree of purity as determined by having an amount of        the total phenolic compound greater than about 80% by weight,        and more preferably greater than about 90% by weight, as        determined by the Folin Ciocalteu method;    -   2. High amount, e.g. between about 25-50% by weight, of low        molecular weight phenolic compounds, wherein low molecular        weight phenolic compounds are monomers, dimers, trimers,        tetramers, and pentamers; and,    -   3. Little to no amount, e.g. less than about 2%, preferably less        than about 1%, of epicatechin-gallate in terminal units and a        small amount, e.g. less than about 12%, preferably less than        about 5%, of epicatechin-gallate in extension units.

Again, while not intending to be bound by any theory, it is believedthat the phenolic profile of grape extracts is important to theireffectiveness in treating or preventing pre-hypertension or MetabolicSyndrome in individuals. In particular, it is believed that the absenceof epicatechin-gallate in terminal units and the small amount ofepicatechin-gallate in extension units of the grape extract used in thepresent invention along with the presence of a higher amount of lowmolecular weight compounds is responsible for increased vasodilatation,which is believed to be responsible for the drop of blood pressure inthe clinical studies of individuals with Metabolic Syndrome andpre-hypertension described below.

Reverse-Phase HPLC Procedure to Determine Percent of Monomers,Oligomers, and Polymers

Reverse-phase HPLC analysis of grape extract can be used to determinethe proportion of monomers, oligomers and polymers based on peak area at280 nm.

HPLC conditions:

Mobile Phase: A: 2% glacial acetic acid B: 80% acetonitrile, 0.4% aceticacid Gradient: Time (min) % A % B Curve  0.00 100  0 —  3.00 100  0 6 6.00  96  4 6  15.00  90 10 6  30.00  85 15 6  50.00  77 23 6  60.00 75 25 6  66.00  70 30 6  80.00  50 50 6  83.00  20 80 6  85.00 100  0 6105.00 100  0 6 110.00 100  0 6 Column: 250 mm × 4.6 mm, Prodigy 5μ ODS(3) 100 Å (Phenomenex, Torrance, CA) Flow rate: 1.0 mL/min Detectionwavelength: 280 nm Temperature: 30° C. Injection: 25 μL

Sample preparation: Accurately weigh 0.1 g grape extract into a 100 mLvolumetric flask. Dissolve the sample in a small amount of methanol (≦5mL), sonicate if necessary. Fill to volume with 18 Megaohm water.Centrifuge the sample (14,000 rpm, 10 min) or filter through 0.45 μMglass filter prior to injection. Determination for percent by weightmonomers, oligomers and polymers is based on the peak area andconcentration of the standards.

Method to Determine Terminal and Extensional Units of ProanthocyanidinsBased on HPLC Analysis After Thiolysis Reaction

Thiolysis is a method to determine average molecular size (degree ofpolymerization) and basic structure of proanthocyanidins in grapeextracts. The information provided may indicate biological quality ofgrape extract for nutritional absorption in the body.

Thiolysis reagent: 5% phenyl methanethiol (benzyl mercaptan) in methanolcontaining 0.2 N HCI.

Condition: 0.1% Grape extract methanol solution was mixed with an equalvolume of thiolysis reagent, stirred, and heated at 90° C. for 2 min.Water was added to stop the reaction. The reactant was then centrifugedat 14000 rpm for 2 min. The supernatant was analyzed directly by HPLC.

HPLC conditions:

Mobile Phase: A: 10% acetic acid/0.1% TFA/5% acetonitrile/84.9% water(v/v/v/v) B: acetonitrile Gradient:  0-30 min  0-50% B 30-35 min 50-100%B Column: 150 cm × 2.0 mm i.d., 4 μm Synergi hydro-RP 80 Å (Phenomenex,Torrance, CA) Flow rate: 0.3 mL/min Detection wavelength: HP 1100 FLDwith excitation @ 276 nm and emission @ 316 nm and HP DAD at 280 nmTemperature: 30° C. Injection: 1-3 μL

The grape extracts to be analyzed were dissolved in methanol, mixed withan equal volume of thiolytic reagent and heated for 2 min at 90° C. Thereleased units were identified by mass spectrometry and quantitativelydetermined by HPLC under the conditions above. The average degree ofpolymerization was measured by calculating the molar ratio of allflavan-3-ol units (thioether adducts plus terminal units) to catechin,epicatechin and epicatechin-gallate corresponding to terminal units. Thepercentage of epicatechin gallate terminal units were determined basedon molar ratio of epicatechin gallate in the sum of total moles ofterminal units, which includes catechin, epicatechin and epicatechingallate. The percentage of epicatechin-gallate extension units weredetermined based on molar ratio of epicatechin gallate thioether adductsin the sum of total moles of thioether adducts of extension units, whichinclude catechin, epicatechin and epicatechin-gallate thioether adducts.The total amount of phenolic compounds was quantified in terms of gramsGallic Acid Equivalents (GAE) by the Folin Ciocalteu method. For moredetails on the Folin Ciocalteu analysis procedure, see: Waterhouse, A.L., Determination of Total Phenolics, in Current Protocols in FoodAnalytical Chemistry, I1.1.1-I1.1.8, Wrolstad, R. E., Wiley, 2001, orSingleton, V. L.; Orthofer, R.; Lamuela-Raventos, R. M. “Analysis oftotal phenols and other oxidation substrates and antioxidants by meansof Folin-Ciocalteu Reagent,” Methods in Enzymology 1999, 299, 152-178,both of which are incorporated herein by reference.

TABLE 1 Comparative Characteristics of MegaNatural ®-BP and Other GrapeExtracts in the Market as Determined by Reverse-Phase HPLC TotalEpicatechin- Epicatechin- Determined by reverse-phase Phenol gallategallate HPLC using peak area g GAE/100 g Terminal Extension MonomersOligomers Polymers Origin Product Name (as is) (%) (%) (%) (%) (%) USAMegaNatural ®-BP 93.8 0.0 6.8 12.8 62.6 24.6 USA MegaNatural ®-BP 91.00.0 5.4 9.2 69.6 21.2 USA MegaNatural ®-BP 95.2 0.0 6.9 13.8 64.7 21.5USA MegaNatural ®-BP 98.6 0.0 8.2 11.3 68.0 20.7 USA MegaNatural ®-BP91.1 0.0 5.4 5.4 71.5 23.2 USA MegaNatural ®-BP 95.5 0.0 5.1 6.5 73.320.3 USA MegaNatural ®-BP 92.7 0.0 3.0 8.5 69.8 21.7 USAMegaNatural ®-BP 93.5 0.0 4.8 5.4 69.9 24.7 USA MegaNatural Gold ® 93.010.5 11.7 7.8 74.7 17.6 USA MegaNatural Gold ® 91.9 4.3 14.6 12.3 76.711.0 USA MegaNatural Gold ® 92.4 11.0 11.9 10.2 77.7 12.1 USAMegaNatural Gold ® 89.1 5.2 12.9 9.9 73.7 16.4 USA MegaNatural Gold ®90.1 3.6 14.5 11.1 73.4 15.6 USA MegaNatural Gold ® 90.3 2.8 8.8 13.265.5 21.3 USA MegaNatural Gold ® 89.6 8.7 11.8 10.0 65.2 24.8 AustraliaVinlife ® 50.6 11.2 11.8 6.3 60.6 33.1 Europe Masquelier OPC ® 98.0 8.57.5 12.1 68.4 19.5 Europe Naturex ® 78.5 8.3 6.2 6.3 64.2 29.5 EuropeIndena ® 93.0 10.5 8.8 10.1 64.4 25.5 China Lycome ® 88.5 10.6 7.5 5.363.1 31.7 China Recovery ® 95.8 3.9 5.4 9.1 58.4 32.5 China Grape P E ®92.6 9.8 6.8 3.6 51.9 44.4 China MA ® 70.1 8.7 10.5 4.6 55.3 40.1 USAME ® 68.9 12.1 6.3 1.9 52.6 45.4 USA San Joaquin ® 74.9 17.7 6.7 2.256.1 41.8 USA Activin ® 84.3 14.8 11.1 3.0 55.9 41.1 Japan KIKKOMAN ®44.5 5.2 13.1 8.1 52.9 38.9Normal-Phase HPLC Analysis for Proanthocyanidins

HPLC analysis of proanthocyanidins: Chromatographic analyses wereperformed on an HP 1100 series HPLC equipped with anautosample/injector, binary pump, column heater, diode array detector,fluorescence detector, and HP ChemStation for data collection andmanipulation. Normal phase separations of proanthocyanidin oligomerswere performed on a Phenomenex Luna Silica (2) column.

Mobile Phase: A: dichloromethane, methanol, water, and acetic acid(83:13:2:2 (v/v)) B: methanol, water, and acetic acid (96:2:2 (v/v))Gradient:  0-30 min linear 0-17.6% B 30-45 min linear 17.6-30.7% B 45-50min linear 30.7-87.8% B 50-60 min linear 87.8% B Column: Phenomenex LUNASilica (3.0 × 150 mm; 3.0 μm) Flow rate: 0.5 mL/min Detection: HP 1100FLD with excitation @ 276 nm and emission @ 316 nm Temperature: 25° C.Injection: 3 μL

In all cases, the column was re-equilibrated between injections withequivalent of 5 mL of the initial mobile phase. Catechin standards wereprepared and analyzed to establish a response calibration curve fromwhich to calculate the concentration of proanthocyanidins in thesamples. Relative response factors of dimers, trimers, tetramers andpentamers to monomers with fluorescence detection were reported by R. L.Prior and L. Gu, “Occurrence and biological significance ofproanthocyanidins in American diet,” Phytochemistry 2005, 66(18)2264-2280, using standards isolated and purified from cocoa bean. Theseresponse factors were used to calculate dimers, trimers, tetramers andpentamers relative to monomers.

TABLE 2 Comparative Characteristics of MegaNatural ®-BP and Other GrapeExtracts in the Market as Determined by Normal-Phase HPLC Determined byNormal-phase HPLC Using Catechin and Epicatechin Equivalent (% byweight) Monomers to Origin Product Name Monomers Dimers TrimersTetramers Pentamers pentamers Others USA MegaNatural ®-BP 6.3 8.7 4.04.4 2.8 26.2 73.8 USA MegaNatural ®-BP 9.1 13.6 5.6 6.2 3.5 38.1 61.9USA MegaNatural ®-BP 9.2 14.8 5.2 5.4 2.7 37.3 62.7 USA MegaNatural ®-BP10.0 14.2 5.4 5.2 2.8 37.6 62.4 USA MegaNatural ®-BP 7.5 11.9 5.2 5.53.4 33.4 66.6 USA MegaNatural ®-BP 8.2 12.1 5.4 5.7 3.5 34.9 65.1 USAMegaNatural ®-BP 9.0 11.3 5.4 4.7 2.5 32.9 67.1 USA MegaNatural ®-BP 5.29.8 4.1 4.8 2.6 26.4 73.6 Australia Vinilife 7.3 4.4 2.0 1.9 1.1 16.783.3 Europe Masquelier OPC ® 15.5 14.2 6.2 4.8 2.9 43.6 56.4 EuropeNaturex ® 8.3 6.4 3.6 3.0 2.0 23.2 76.8 Europe Indena ® 16.6 12.4 6.24.8 3.3 43.3 56.7 China Lycome ® 7.6 6.2 3.4 2.8 1.9 22.0 78.0 ChinaRecoverye ® 17.5 8.1 4.4 2.8 2.0 34.8 65.2 China Grape P E ® 3.9 3.2 1.91.5 1.1 11.6 88.4 China MA ® 4.1 3.8 1.9 1.7 0.8 12.3 87.7 USA ME ® 1.81.8 0.9 0.8 0.5 5.8 94.2 USA San Joaquin ® 5.3 5.8 2.6 1.8 1.3 16.8 83.2USA Activin ® 5.5 4.8 2.1 1.3 1.3 15.1 84.9 Japan KiKKOMAN ® 0.9 1.1 0.70.7 0.4 3.7 96.3

The results as set forth in Tables 1 and 2 were obtained using differentmethods, which account for the different ranges of, for example, thepercent of monomers. For instance, reverse-phase HPLC was used todetermine percentage of monomers, oligomers and polymers based on peakareas of these three groups of compounds. Gallaic acid is included asmonomers. In the normal phase HPLC, catechin and epicatechin were usedas standards to determine the amount of monomers, dimers, trimers,tetramers and pentamers in the grape extract by weight. The relativeresponse factors of dimers, trimers, tetramers and pentamers related tomonomers reported by R. L. Prior and L. Gu were used to calculatedimers, trimers, tetramers and pentamers.

2. Effect of Grape Extract on Blood Pressure of Individuals withMetabolic Syndrome

The effects of grape extract used in the present invention on bloodpressure were studied in twenty-four individuals diagnosed withMetabolic Syndrome. The study included an equal number of men and womenbetween the ages of 20 and 50 years. Metabolic Syndrome was diagnosed onthe basis of the criteria defined by the National Cholesterol EducationProgram Adult Treatment Panel III. Each subject exhibited at least threeof the following features: 1) Fasting blood sugar >110 mg/dL, 2) HDL(<40 mg/dL in men and <45 mg/dL in women), 3) blood pressure >130/85,and 4) abdominal obesity (>102 cm for men and >88 cm for women).Individuals were excluded if they were current smokers or ex-smokers (<3years); taking anti-inflammatory or hypertensive drugs; or consumedover-the-counter anti-oxidant compounds.

The individuals were block randomized into three groups of eight andthey were given one of the following capsules depending on their groupassignment.

-   -   Group 1 was given a placebo capsule    -   Group 2 was given a capsule containing 150 mg Grape Extract    -   Group 3 was given a capsule containing 300 mg Grape Extract

The individuals were given sufficient capsules to take the same doseonce daily for the next twenty-eight days. At the end of this period,blood pressure measurements were obtained. Ambulatory blood pressureswere recorded over a 12 hr period at the commencement of the study andagain after four weeks. The procedure was non-invasive and involvedplacing a blood pressure cuff in the upper arm. The cuff was connectedto an automated FDA-approved inflating device, which was worn on a belt.

Table 3 shows the blood pressure data for the three groups ofindividuals with Metabolic Syndrome. In those receiving 300 mg and 150mg daily of the grape extract used in the present invention, there weresignificant reductions in both systolic and diastolic pressure. Therewere no significant changes in the group given placebo.

TABLE 3 Results of Grape Extract Used in the Present Invention on BloodPressure of Individual with Metabolic Syndrome 300 mg daily 150 mg dailyPlacebo Systolic Diastolic Systolic Diastolic Systolic Diastolic Start129 ± 4 79 ± 3 137 ± 4 84 ± 3.3 124 ± 4 74 ± 4 4 weeks 117 ± 3 71 ± 3125 ± 4 78 ± 1.9 123 ± 4 71 ± 4 p* 0.007 0.006 0.003 0.009 ns ns *p isthe probability that the start and end values are the same. A p of 0.05or less (5%) is generally considered significant.

The relationship between the baseline blood pressure and the fall inboth systolic and diastolic pressures are shown in FIGS. 1 and 2. Bloodpressure is quoted as mmHg. Since the diagnosis of Metabolic Syndrome isbased on the presence of three of the listed risk factors (one of whichis blood pressure), the study did not block randomize the individualsfor blood pressure. As such, the average pressures in the three groupswere not similar (but varied within a narrow range).

This study demonstrates that the grape extract of the present inventionin daily dosage of 150 mg and 300 mg lowers both systolic and diastolicblood pressure in individuals with Metabolic Syndrome. The fall in bloodpressure is statistically significant for both doses of extract used. Infact, the changes in blood pressure observed with using grape extractwere comparable to those observed in major clinical trials usingpharmaceutical agents.

3. Effect of Grape Extract on Oxidized LDL of Individuals with MetabolicSyndrome

The effects of grape extract used in the present invention on oxidizedLDL were studied in the same twenty-four individuals diagnosed withMetabolic Syndrome as discussed above. The concentration of oxidized LDLwas measured at the commencement of the study and again after four weeksof treatment. In order to measure the concentration of oxidized LDL, ablood sample was taken from each individual and analyzed.

The changes in concentration of oxidized LDL for the three groups aresummarized in FIG. 3. FIG. 3 shows a slight drop in oxidized LDL forplacebo, a trend drop in oxidized LDL for individuals taking 150 mg ofthe grape extract used in the present invention, and a statisticallysignificant drop (p<0.05) in oxidized LDL for individuals taking 300 mgof the grape extract used in the present invention. FIG. 4 depicts therelationship between the change in oxidized LDL and the baselineconcentration of oxidized LDL in individuals given 300 mg of the grapeextract used in the study. The regression coefficient, R², =0.52. FIG. 4shows a greater drop in oxidized LDL concentration in individuals whostarted with higher levels of oxidized LDL to begin with.

This study demonstrates that the grape extract of the present inventionin daily dosage of 150 mg and 300 mg reduces oxidized LDL concentrationin plasma in individuals with Metabolic Syndrome. Further, there was astatistically significant decrease in oxidized LDL concentration forthose individuals given 300 mg of the grape extract used in the presentinvention.

4. Effect of Grape Extract on Individuals with Pre-Hypertension

The effects of grape extract used in the present invention were studiedon twenty-four individuals diagnosed with pre-hypertension. The studyincluded an equal number of men and women between the ages of 30 and 60years. Pre-hypertension was diagnosed on the basis of the criteriadefined by the Seventh Report of the Joint National Committee onPrevention, Detection, Evaluation, and Treatment of High Blood Pressure.Each subject had systolic pressure between 120 and 139 mmHg and/ordiastolic pressure between 81 and 89 mmHG. Individuals were excluded ifthey were current smokers or ex-smokers (<3 years); takinganti-inflammatory or hypertensive drugs; or consumed over-the-counteranti-oxidant compounds.

The individuals were block randomized for gender into two groups oftwelve and they were given one of the following capsules depending ontheir group assignment.

-   -   Group 1 was given a placebo capsule    -   Group 2 was given a capsule containing 300 mg MegaNatural®-BP

The individuals were given sufficient capsules to take the same doseonce daily for the next eight weeks. At the end of this period, bloodpressure measurements were obtained. Ambulatory blood pressures wererecorded over a 12 hr period at the commencement of the study and againafter eight weeks. The procedure was non-invasive and involved placing ablood pressure cuff in the upper arm. The cuff was connected to anautomated FDA-approved inflating device, which was worn on a belt.

Table 4 shows the blood pressure data for the two groups of individualswith pre -hypertension. The baseline pressures between the two groupswere not significantly different. In those receiving 300 mg daily of thegrape extract used in the present invention, there were significantreductions in both systolic and diastolic pressure; however, there wereno significant changes in the group given placebo. For instance, theaverage drop in systolic blood pressure in the group treated withMegaNatural®-BP was 7.2±2.5 mmHg, whereas the systolic blood pressure inthe placebo group increased by 0.03±1.5 mmHg. The data is summarizedbelow. The values are given in mmHg (mean±SEM).

TABLE 4 Results of MegaNatural ®-BP on Blood Pressure of Individual withPre-Hypertension 300 mg MegaNatural ®- BP daily Placebo SystolicDiastolic Systolic Diastolic Start 133 ± 2 80 ± 2 134 ± 2 79 ± 2 8 weeks126 ± 2 73 ± 2 134 ± 2 80 ± 2 p* 0.021 0.042 ns ns *p is the probabilitythat the start and end values are the same. A p of 0.05 or less (5%) isgenerally considered significant.

This study demonstrates that the grape extract of the present inventionin daily dosage of 300 mg lowers both systolic and diastolic bloodpressure in individuals with pre-hypertension. The fall in bloodpressure is statistically significant. In fact, the changes in bloodpressure observed with using grape extract were comparable to thoseobserved in major clinical trials using pharmaceutical agents.

EXAMPLES

The invention is further defined by reference to the following examplesdescribing a process for making the grape extract and preparing thedietary supplements. The examples are representative, and they shouldnot be construed to limit the scope of the invention.

Example 1 Process for Making the Grape Extract

Dried grape seeds were extracted with water at a temperature of 200° F.for two hours and the extract was separated from the seeds on metalscreens. The extract was cooled to 90-100° F. and pectinase was added ata concentration of 200 ppm. The extract was divided into two portions.To one portion, commercial fungal tannase enzyme (tannin acylhydrolase,E.C3.1.1.20) was added at a concentration of 1000 ppm. To the secondportion, the tannase was added at a concentration of 50 ppm. Theresidual concentration of gallic acid in the original extract was 117ppm with 18.9% terminal units and 11.1% extension units. Within abouttwo hours of treatment with 1000 ppm tannase enzyme, the gallic acidconcentration rose to 904 ppm with 0% terminal units and about 5.5%extension units. It took about thirty-four hours of treatment with 50ppm tannase enzyme for the gallic acid to reach 810 ppm with less than1% terminal units and less than 6% extension units. After about twodays, both the extracts were acidified to a pH of 1.5 to 2.5, whichallowed flocculation of proteins and polysaccharides on cooler storagefrom 40-60° F. The extract was filtered and processed further accordingto the '581 patent to produce a grape extract with characteristics forblood pressure reduction and reduction of oxidized LDL concentrations.

Example 2 Capsules

MegaNatural®-BP grape extract (150 mg or 300 mg) was dry mixed withmagnesium stearate (3 mg or 6 mg respectively) and loaded into hardshell gelatin capsules (made of gelatin and water). In the 150 mgformulation, the grape extract has a minimum of 90% phenols or 135 mg ofphenols per 150 mg of grape extract. In the 300 mg formulation, thegrape extract has a minimum of 90% phenols or 270 mg of phenols per 300mg of grape extract. The daily dosage is one capsule per day.

Example 3 Powder

MegaNatural®-BP grape extract was formulated into a dry mix with theexcipients as shown in Table 4 to be used in a beverage, wherein theingredients were dry blended. To prepare the final beverage, 9.47 g ofthe dry mix is combined with 500 mL of cold water and stirred. A 500 mLserving contains 16 calories. The final beverage contains 100 mgMegaNatural®-BP grape extract and 120 mg vitamin C per 1 L serving,which will have an ORAC value of 2200 TE.

ORAC, measured in mmoles Trolox (a noncommercial, water-solublederivative of tocopherol) equivalents (TE) per gram, stands for “OxygenRadical Absorbance Capacity.” This is the standard by which scientistsmeasure antioxidant activity in foods and supplements. A single servingsof fresh or freshly cooked fruits and vegetables supply an average of600 to 800 ORAC units. It has been suggested that increasing intake offoods or supplements that provide 2,000 to 5,000 ORAC units per day mayhave health benefits.

TABLE 4 Ingredients % Dry Mix (g) Maltodextrin 37.48 Citric Acid 29.99Clouding Agent (Purity Gum 2000)* 5.25 Aspartame 3.85 Sodium Citrate,FCC Grade 3.75 Ultra Guar** 3.75 N&A Orange Flavor (SN313897)*** 7.5 NatFF Passion Fruit Flavor (SN 313898)*** 4.27 FD&C Yellow #6(20:1 inMaltodextrin) 2.24 FD&C Yellow #5(20:1 in Maltodextrin) 0.75 AscorbicAcid 0.64 MegaNatural ®-BP Grape Extract (Polyphenolics, Inc.) 0.53TOTAL 100 *Available from National Starch & Chemical Corporation,Bridgewater, NJ **Available From P. L. Thomas & Co., Inc. Morristown, NJ***Available from International Flavors & Fragrances, Dayton, NJ

Example 4 Beverage

MegaNatural®-BP grape extract was formulated into a beverage with theexcipients as shown in Table 5. The following beverage contains 50 mgMegaNatural®-BP grape extract and 60 mg vitamin C (100% RDI) per 8 fl ozserving. In an 8 fl oz serving, the beverage contains 0 calories and0.15 g total carbohydrates. A 16 fl oz serving would have an ORAC valueof 2200 TE.

TABLE 5 Ingredients % by weight Water 99.4373 Citric Acid 0.2640MegaNatural Purple Color (Canandaigua Conc.) 0.0528 Sethness - GreenleafFlavor System 0.0867 Sodium Benzoate 0.0448 Potassium Sorbate 0.0448Ascorbic Acid 0.0338 MegaNatural ®-BP Grape Extract (Polyphenolics,Inc.) 0.0211 Asparatame 0.0147 TOTAL 100.0000%

Example 5 Beverage

MegaNatural®-BP grape extract was formulated into a beverage with theexcipients as shown in Table 6. The following beverage contains 50 mgMegaNatural®-BP grape extract and 60 mg vitamin C (100% RDI) per 8 fl ozserving. In an 8 fl oz serving, the beverage contains 15 calories and 4g total carbohydrates. A 16 fl oz serving would have an ORAC value of2200 TE.

TABLE 6 Ingredients % by weight Water 95.8778 Orange Juice Concentrate65 1.3973 Cranberry Juice Concentrate 50 0.8691 MegaNatural Purple Color(Canandaigua Conc.) 0.5032 Sethness - Greenleaf Flavor System 1.1074Sodium Benzoate 0.0444 Potassium Sorbate 0.0444 Ascorbic Acid 0.0357MegaNatural ®-BP Grape Extract (Polyphenolics, Inc.) 0.0210 Neotame (TheNutraSweet Co.) 0.0997 TOTAL 100.0000%

Example 6 Vitamin/Mineral Supplement

MegaNatura®-BP grape extract (150 mg) was dry mixed with the followingexcipients listed in Table 7 and pressed into a tablet to form amulti-vitamin/mineral supplement. The daily dosage is one tablet perday, preferably taken with food.

TABLE 7 Ingredients % Daily Value Vitamin A 3500 IU (29% as BetaCarotene) 70 Vitamin C 60 mg 100 Vitamin D 400 IU 100 Vitamin E 45 IU150 Vitamin K 10 mcg 13 Thiamin 1.5 mg 100 Riboflavin 1.7 mg 100 Niacin20 mg 100 Vitamin B6 3 mg 150 Folic Acid 400 mcg 100 Vitamin B12 25 mcg417 Biotin 30 mcg 10 Pantothenic Acid 10 mg 100 Calcium 299 mg 20Phosphorus 48 mg 5 Iodine 150 mcg 100 Magnesium 100 mg 25 Zinc 15 mg 100Selenium 20 mg 29 Copper 2 mg 100 Manganese 2 mg 100 Chromium 150 mcg125 Molydenum 75 mcg 100 Chloride 72 mg 2 Potassium 80 mg 2MegaNatural ®-BP Grape Extract 150 mg * Boron 150 mcg * Nickel 5 mcg *Silicon 2 mg * Vanadium 10 mcg * Lutein 250 mcg * Lycopene 300 mcg **Daily Value (% DV) not established

Example 7 Vitamin/Mineral Supplement

MegaNatural®-BP grape extract (150 mg) was blended with the followingingredients and excipients listed in Table 8 in V blender until uniform.The blend was pressed into tablets that reach a specified weight of 775mg±2% to form a multi-vitamin/mineral supplement. The tablets were spraycoated with a clear coating of a water soluble gum such as hydroxypropylmethylcellulose and dried. The daily dosage is one tablet per day. Thebatch size for the formulation in Table 8 is 500,000 Tablets.

TABLE 8 Amount/ Amount/ Label Overage Tablet Batch Ingredients (Units ofMeasure) Claim (%)* (mg) (Kg) Vitamin A Palmitate @ 500 K IU/gm (IU)5000 IU 30 13.000 6.500 Vitamin D₃ @ 850K IU/g (IU) 400 IU 30 0.6120.306 Vitamin E succinate (D-α) @ 1210 IU/g (IU) 15 IU 5 13.017 6.508Vitamin C (mg) 30 mg 2 30.600 15.300 Thiamine HCl @ 89.2% (mg) 1.5 mg 21.715 0.858 Riboflavin (mg) 1.7 mg 2 1.734 0.867 Niacinamide (mg) 10 mg2 10.200 5.100 Pyridoxine HCl 82.3% (mg) 2 mg 5 2.552 1.276 Folic AcidTrituration 1.0% (mcg) 400 mcg 25 50.000 25.000 Vitamin B-12 Trituration1.0% (mcg) 6 mcg 20 0.720 0.360 Pantothenic Acid (Cal Pan.) (mg) 10 mg 510.500 5.250 Biotin Trituration 1.0% (mcg) 30 mcg 20 3.600 1.800 Calcium(Dicalcium Phosphate) 29.46% (mg) 100 mg 0 344.119 172.060 Phosphorus(Dicalcium Phosphate) 22.77% (mg) 75 mg 0 0.000 0.000 Magnesium (MgO)60.32% (mg) 20 mg 0 33.156 16.578 Zinc (ZnO) 80.34 (mg) 5 mg 0 6.2243.112 Iodine (KI) 76.45% (mcg) 150 mcg 0 0.196 0.098 Copper (Gluconate)14.00% (mg) 2 mg 0 14.286 7.143 Manganese (Gluconate) 12.34% (mg) 2 mg 016.207 8.104 MegaNatural ®-BP Grape Extract 150 mg 150.000 25.000Microcrystalline cellulose 33.750 16.875 Croscarmellose Sodium 20.25010.125 Stearic Acid 13.500 6.750 Magnesium Stearate 5.063 2.531 TOTAL775.000 337.500 *Percent amount of ingredient over label claim used toreach the label claim amount.

1. A method of treating Metabolic Syndrome in a mammal in need thereof,comprising administering to the mammal a composition comprising apolyphenol extract from grapes in an amount effective to lower bloodpressure, wherein the extract comprises about 2% by weight or lessepicatechin-gallate terminal units, and monomers, dimers, trimers,tetramers and pentamers in a total amount between about 25-50% by weightof the extract.
 2. The method of claim 1, wherein the extract comprisesabout 80% by weight or more total phenolic compounds.
 3. The method ofclaim 2, wherein the extract comprises about 12% by weight or lessepicatechin-gallate extension units.
 4. The method of claim 1, whereinthe amount of extract in the composition is between about 50-1000 mg. 5.The method of claim 1, wherein the composition is formulated for oraladministration.
 6. The method of claim 5, wherein the composition isadministered in a form selected from the group consisting of tablets,powders, liquids, capsules and gels.
 7. The method of claim 5, whereinthe composition is a food or a beverage product.
 8. The method of claim5, wherein the composition is a nutraceutical.
 9. The method of claim 1,wherein the mammal is a human.
 10. The method of claim 1, wherein theextract contains 400-1500 ppm of gallic acid.
 11. A method of treatingpre-hypertension in a mammal in need thereof, comprising administeringto the mammal a composition comprising a polyphenol extract from grapesin an amount effective to lower blood pressure, wherein the extractcomprises about 2% by weight or less epicatechin-gallate terminal units,and wherein the total amount of monomers, dimers, trimers, tetramers andpentamers is between about 25- 50% by weight.
 12. The method of claim11, wherein the extract comprises about 80% by weight or more totalphenolic compounds.
 13. The method of claim 12, wherein the extractcomprises about 12% by weight or less epicatechin-gallate extensionunits.
 14. The method of claim 11, wherein the amount of extract in thecomposition is between about 50-1000 mg.
 15. The method of claim 11,wherein the composition is formulated for oral administration.
 16. Themethod of claim 15, wherein the composition is administered in a formselected from the group consisting of tablets, powders, liquids,capsules and gels.
 17. The method of claim 15, wherein the compositionis a food or a beverage product.
 18. The method of claim 15, wherein thecomposition is a nutraceutical.
 19. The method of claim 11, wherein themammal is a human.
 20. The method of claim 11, wherein the extractcontains 400-1500 ppm of gallic acid.
 21. A method of treating MetabolicSyndrome in a mammal in need thereof, comprising administering to themammal a composition comprising a polyphenol extract from grapes in anamount effective to reduce oxidized LDL cholesterol, wherein the extractcomprises about 2% by weight or less epicatechin-gallate terminal units,and wherein the total amount of monomers, dimers, trimers, tetramers andpentamers is between about 25-50% by weight.
 22. The method of claim 21,wherein the extract comprises about 80% by weight or more total phenoliccompounds.
 23. The method of claim 22, wherein the extract comprisesabout 12% by weight or less epicatechin-gallate extension units.
 24. Themethod of claim 21, wherein the amount of extract in the composition isbetween about 50-1000 mg.
 25. The method of claim 21, wherein thecomposition is formulated for oral administration.
 26. The method ofclaim 25, wherein the composition is administered in a form selectedfrom the group consisting of tablets, powders, liquids, capsules andgels.
 27. The method of claim 25, wherein the composition is a food or abeverage product.
 28. The method of claim 25, wherein the composition isa nutraceutical.
 29. The method of claim 21, wherein the mammal is ahuman.
 30. The method of claim 21, wherein the extract contains 400-1500ppm of gallic acid.
 31. A method of treating Metabolic Syndrome in amammal in need thereof, comprising administering to the mammal acomposition comprising a polyphenol extract from grapes having about5-15% monomers, about 5-20% dimers, about 3-10% trimers, about 2-10%tetramers, and about 2-10% pentamers by weight, wherein the extractcomprises about 2% by weight or less epicatechin-gallate terminal units.32. The method of claim 31, wherein the extract contains 400-1500 ppm ofgallic acid.
 33. A method of treating pre-hypertension in a mammal inneed thereof, comprising administering to the mammal a compositioncomprising a polyphenol extract from grapes having about 5-15% monomers,about 5-20% dimers, about 3-10% trimers, about 2-10% tetramers, andabout 2-10% pentamers by weight, wherein the extract comprises about 2%by weight or less epicatechin-gallate terminal units.
 34. The method ofclaim 33, wherein the extract contains 400-1500 ppm of gallic acid. 35.A method of treating Metabolic Syndrome in a mammal in need thereof,comprising administering to the mammal a composition comprising apolyphenol extract from grapes in an amount effective to lower bloodpressure, wherein the extract comprises about 2% by weight or lessepicatechin-gallate terminal units, and between about 5-15% monomers,about 5-20% dimers, about 3-10% trimers, about 2-10% tetramers, andabout 2-10% pentamers by weight.